The goal of this project is to provide facilities such as public health laboratories or hospital emergency rooms with a practical, fully validated immunodiagnostic platform and multiplexed immunoassays that can be used for the simultaneous detection of multiple biothreat agents and to distinguish pathogens that cause common respiratory infections from biothreat agents that cause infections after exposure. Using Luminex-based liquid arrays, we have developed and demonstrated multiplexed immunoassays that can be used for the simultaneous detection of 11 different biothreat agents from a single sample. Multiplexed immunoassays designed for detection of respiratory pathogens will be developed and tested in a clinical environment. By combining multiplexed assays for detection of respiratory pathogens with multiplexed assays for detection of biothreat agents, a single sample can be screened for a large number of agents while simultaneously distinguishing biothreat agents from common respiratory pathogens. To find wide acceptance in the user community, we will validate the platform and our multiplexed immunoassays, and make the validated assays commercially available. Sample processing protocols will be developed to ensure maximum sensitivity, specificity, and selectivity from clinical samples comprising complex sample matrices. Assay performance in clinical samples will be optimized to provide a diagnostic tool that performs better than the currently employed clinical gold standards. Using multivariate statistical analysis, we will exploit the power of multiplexed analysis to develop organism-specific signature profiles that can be used to further distinguish true positives in potential-confounding sample matrices. Researchers from LLNL and the UCDavis Medical Center, in collaboration with the CDC, have partnered with Tetracore, Inc., Bio-Rad, and Chemicon International to form a very powerful, multidisciplinary team with the collective resources, expertise and motivation to achieve these goals. This work will result in commercially available, fully validated, multiplexed immunoassays and an immunodiagnostic platform that is rapid, sensitive, specific, cost-effective, and capable of simultaneously detecting multiple biological threat agents. [unreadable] [unreadable]